Title Page
ABSTRACT
국문 초록
PREFACE
Contents
ABBREVIATION 16
CHAPTER 1. INTRODUCTION 19
1.1. Ambient air pollution is a major factor of pneumonitis, associated with significant mortality patients with COPD 19
1.2. Acrolein, one of air pollutant components, induces acute intracellular GSH depletion 20
1.3. Curcumin enhances intracellular GSH level through interaction with Nrf2-Keap1 signaling 20
CHAPTER 2. RESULTS 22
2.1. Curcumin reduces pneumonitis-related circulating inflammatory cytokines in subjects in Chengdu City 22
2.2. Curcumin reduces acrolein-induced pneumonitis by suppressing DAMPs release through apoptotic alveolar epithelial cell death 28
2.3. Curcumin suppresses intrinsic and extrinsic apoptotic signaling pathways triggered in the response to acrolein 38
2.4. Curcumin inhibits DNA damage response and JNK/c-Jun signaling pathway in a ROS-dependent manner 46
2.5. Curcumin inhibits DNA damage response and JNK/c-Jun signaling pathway in a ROS-dependent manner 56
CHAPTER 3. DISCUSSION 63
CHAPTER 4. CONCLUSION 65
REFERENCES 66
APPENDIX (SUPPLEMENTARY MATERIALS) 75
Table 1. Baseline characteristics and immune-related markers in plasma of subjects 25
Figure 1. Human serum analysis for circulating inflammatory cytokines 26
Figure 2. Circulating cytokine ratio in human serum for inflammatory response analysis 27
Figure 3. Measuring circulating pro-inflammatory cytokines in mice serum with ELISA assay 31
Figure 4. Histological phenotype assessment in acrolein-inhaled mice lung tissue (x20) 32
Figure 5. Immunofluorescence image observation for fibrotic pathogenesis-related markers (x40) 33
Figure 6. Characteristics of infiltrating immune cells in lung tissues by immunofluorescence (x40) 34
Figure 7. HMGB1, one of DAMP signal molecules, detected under fluorescence image counterstained with DAPI 35
Figure 8. Pattern of ICD, especially pyroptosis (N-GSDMD) and necroptosis (p-MLKL) 36
Figure 9. TUNEL, the cell death marker in alveolar epithelial cells counterstained with DAPI (x40) 37
Figure 10. Immunofluorescence image for c-caspase 3 expression analysis counterstained with SpC 40
Figure 11. Immunoblot data for analysis apoptosis-related markers 41
Figure 12. Immunostained in lung tissue for intrinsic apoptosis signaling marker, c-caspase 9 42
Figure 13. Protein expression for intrinsic apoptosis-related markers 43
Figure 14. Protein expression for extrinsic apoptosis-related markers 44
Figure 15. Immunofluorescence analysis for intrinsic and extrinsic apoptosis axis with spatial distribution of Bax and c-caspase 8 45
Figure 16. p53 protein localization and expression analysis 48
Figure 17. Immunofluorescence analysis for p-ATM, known as critical player in p53 activation 49
Figure 18. Protein expression and spatial distribution analysis for γH2AX 50
Figure 19. Immunofluorescence analysis for c-Jun, the H2AX phosphorylation inducer, with spatial distribution 51
Figure 20. Immunofluorescence analysis for p-JNK, promoter of apoptosis that sensitized by oxidative stress 52
Figure 21. Immunofluorescence analysis for 8-OHdG, representative marker for DNA oxidation 53
Figure 22. Immunofluorescence analysis for 4-HNE, representative marker for lipid oxidation 54
Figure 23. Total GSH expression analysis using GSH adduct antibody 55
Figure 24. Remained acrolein examination with acrolein adduct antibody 58
Figure 25. Protein expression for GSH conjugation-related enzyme, GST 59
Figure 26. Protein expression for GSH synthesis-related enzymes, GSS, GCLM, and GCLC 60
Figure 27. Nrf2 protein expression (total Nrf2) and localization (activated Nrf2) analysis 61
Figure 28. Examination on Keap1 protein degradation using immunostained image 62